Abstract
Recently developed single-cell RNA sequencing methods allow the simultaneous profiling of the transcriptomes of thousands of individual cells. However, current methods still require advanced equipment or entail substantial waste of reagents. Here, we introduce magnetic bead-assisted parallel single-cell gene expression sequencing (MAPS-seq), a microwell-based method that pools samples before the reverse transcription step, increasing the ease of sample preparation and reducing reagent waste. Moreover, because this method uses universal reagents and standard molecular biology lab instruments, it is easy to implement, even in labs that have not previously conducted single-cell RNA sequencing. We validated our method by demonstrating that it can generate gene expression data at the single-cell level. We then applied the MAPS-seq method to analyze 237 human myelogenous leukemia cells treated with one of three different drugs or dimethyl sulfoxide. We observed transcriptional changes and identified marker genes that indicate a drug response. Furthermore, the MAPS-seq method produced data of comparable quality to those of existing single-cell RNA sequencing methods. Consequently, we expect that our method will provide researchers with a more accessible, less wasteful, and less burdensome method for investigating the transcriptomes of individual cells.
Original language | English |
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Pages (from-to) | 804-814 |
Number of pages | 11 |
Journal | Experimental and Molecular Medicine |
Volume | 52 |
Issue number | 5 |
DOIs | |
Publication status | Published - 2020 May 1 |
Bibliographical note
Funding Information:This work was supported by the following sources: (i) the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT and Future Planning (grant no. NRF-2018R1A2B2001322); (ii) the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI) funded by the Ministry of Health & Welfare, Republic of Korea (grant no. HI18C2282); (iii) the Bio & Medical Technology Development Program of the National Research Foundation (NRF) funded by the Korean government (MSIT) (No. 2016M3A9B6026918); (iv) the Mid-career Researcher Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT and Future Planning (grant no. NRF-2018R1A2A1A05079172); and (v) the Bio & Medical Technology Development Program of the National Research Foundation (NRF) funded by the Korean government (grant no. NRF-2016M3A9B6948494).
Publisher Copyright:
© 2020, The Author(s).
All Science Journal Classification (ASJC) codes
- Biochemistry
- Molecular Medicine
- Molecular Biology
- Clinical Biochemistry